Figure 5

SH2B1β enhances the phosphorylation levels of AKT, ERK1/2 and FoxOs. PC12-GFP and PC12-SH2B1β cells were incubated in serum-free medium overnight followed by H₂O₂ stimulation with the indicated concentrations for 10 min. Equal amounts of proteins from the lysates were resolved via SDS-PAGE and immunoblotted with (A) anti-GFP; (B) anti-pAKT(Ser473) and anti-AKT; (D) anti-pERK1/2 and anti-ERK1/2; (F) anti-FoxO1 and anti-FoxO3a antibodies. Representative blots were shown. (C) The quantified results of pAKT in PC12-GFP and PC12-SH2B1β cells were shown. The levels of pAKT were normalized to levels of AKT at each condition. The relative level of pAKT at 50 μM of H₂O₂ in PC12-SH2B1β cells was defined as 100% in each experiment and others were normalized to this value. (E) The levels of pERK1/2 were normalized to total ERK1/2 levels, the relative level of pERK1/2 at 300 μM in PC12-SH2B1β cells was defined as 100% for each experiment and others were normalized to this value. Data are expressed as mean ± S.E. from three (for AKT) or five (for ERK1/2) independent experiments. Arrows point to the phospho-FoxO1 and 3a.